JP4917329B2 - Image acquisition apparatus, image acquisition method, and image acquisition program - Google Patents

Abstract

A semi-automatic mode according to which an operator makes a necessary confirmation when an image pickup condition for a micro image is set with reference to a macro image is provided in controlling the acquisition of the macro image, the setting of the image pickup condition, and the acquisition of the micro image with respect to each of a plurality of samples (S) stored in a sample storage unit (11), and in the semi-automatic mode, the sample (S) whose macro image has been acquired by a macro image acquiring unit (20) is placed at a standby position without being moved directly to an image acquiring position for a micro image acquiring unit (30). According to this structure, image acquisition with respect to each of the samples (S) can be efficiently performed, and an operational burden imposed on the operator is reduced. Accordingly, it becomes possible to realize an image acquiring apparatus, an image acquiring method, and an image acquiring program each of which is capable of performing image acquisition with respect to each of the samples with high efficiency.

Description

  The present invention relates to an image acquisition apparatus, an image acquisition method, and an image acquisition program for acquiring an image of a sample.

  In recent years, in the field of pathology and the like, a virtual microscope that can be operated as if a sample is being viewed with an actual microscope in a virtual space such as a personal computer is known. The sample data handled by this virtual microscope is based on image data of a sample acquired in advance with high resolution using an actual microscope.

As described above, an image acquisition apparatus that acquires image data of a sample used in a virtual microscope is required to acquire an image of the sample with a sufficiently high resolution in order to realize an image operation with the virtual microscope. In order to efficiently acquire such a high-resolution image, a configuration in which a sample image is acquired using a macro image acquisition camera and a high-resolution micro image acquisition camera is disclosed in Patent Document 1. : Described in US Pat. No. 6,816,606.
US Pat. No. 6,816,606

  In the configuration including the macro camera and the micro camera as described above, for example, the macro image is first acquired by the macro camera, the imaging condition for the sample is set with reference to the macro image, and then, A method of acquiring a micro image with a micro camera with reference to set imaging conditions is conceivable. In addition, when acquiring image data of a sample used in a virtual microscope, a large number of slides in which a biological sample or the like is sealed on a slide glass are prepared as samples, and image acquisition is continuously performed for the plurality of slides. There is a case. In such a case, in the above configuration, the macro image and micro image acquisition operations are repeatedly performed on the sample.

  In such image acquisition of a plurality of samples, if the imaging conditions using the macro image are automatically set, the acquisition of the macro image, the setting of the imaging conditions, and the acquisition of the micro image for each sample are automatically performed sequentially. Is possible. However, in such a method, for example, if there is dust other than a biological sample in the slide, the imaging conditions for the slide may not be set correctly, and a normal micro image may not be acquired. On the other hand, in the method of manually setting the imaging conditions for each sample, the operator is completely attached to the image acquisition device until the image acquisition for each of the plurality of samples is completed, and the operator's work load is reduced. growing.

  The present invention has been made to solve the above problems, and provides an image acquisition apparatus, an image acquisition method, and an image acquisition program capable of efficiently performing image acquisition for each of a plurality of samples. The purpose is to provide.

In order to achieve such an object, an image acquisition device according to the present invention includes (1) a sample storage means for storing a plurality of samples, (2) a macro image acquisition means for acquiring a macro image of the sample, (3) Micro image acquisition means for scanning the sample at a higher resolution than the macro image and acquiring the micro image; and (4) Macro image acquisition control for controlling the macro image acquisition operation by the macro image acquisition means. comprising means, image pickup condition setting means for setting the imaging conditions of the reference to micro image the macro image, and a control unit having a micro image acquisition control means for controlling an operation to acquire the micro image by the micro image acquiring means, image pickup condition The setting means refers to the macro image, sets an image acquisition range and a focus measurement position as imaging conditions for the micro image, and the micro image acquisition control means sets the focus measurement position to the focus measurement position. Acquires focus information for the sample by Zui performs control to acquire the micro image based on the focus information and the image acquiring range obtained, (5) the control means has a semi-automatic mode serving as a control mode, ( 6) In the semi-automatic mode, the macro image acquisition control unit arranges the sample at the storage position by the sample storage unit at each image acquisition position by the macro image acquisition unit for each of the plurality of samples, Furthermore, control is performed to place the sample at the standby position, and the imaging condition setting means sets the imaging condition for the micro image by allowing the operator to check the macro image and the corresponding imaging condition for the sample from which the macro image has been acquired. as well as, after the macro image of the first sample has been obtained, in parallel with the acquisition of the macro image, to set the image pickup condition, the micro image The resulting control means performs control to an imaging condition In the standby position is set samples were placed at an image acquiring position for the micro image acquiring means, to acquire the micro image with reference to the image pickup condition, the semi-automatic mode The imaging condition setting means displays the imaging condition automatically obtained corresponding to the macro image on the display means, and selects whether to use the imaging condition or to manually reset the imaging condition. It is characterized by making it.

The image acquisition method according to the present invention includes (1) a plurality of samples stored in the sample storage means, (2) a macro image acquisition step for acquiring a macro image of the sample, and (3) a macro image of the sample. A micro image acquisition step for scanning the image at a higher resolution and acquiring the micro image, and (4) a macro image acquisition control step for controlling the macro image acquisition operation by the macro image acquisition step. An imaging condition setting step for setting an imaging condition for the image, and a control step having a micro image acquisition control step for controlling a micro image acquisition operation by the micro image acquisition step. The imaging condition setting step refers to a macro image. Then, an image acquisition range and a focus measurement position are set as imaging conditions for the micro image, and the micro image acquisition control step Acquires focus information for the sample based on the measurement position, performs control to acquire the micro image based on the focus information and the image acquiring range obtained, (5) control step, have a semi-automatic mode serving as a control mode (6) In the semi-automatic mode, in the macro image acquisition control step, for each of the plurality of samples, the sample storage unit places the sample in the storage position at the image acquisition position in the macro image acquisition step, and displays the macro image. The imaging condition setting step allows the operator to check the macro image and the corresponding imaging conditions for the sample from which the macro image has been acquired. sets a condition, after the macro image of the first sample has been obtained, in parallel with the acquisition of the macro image, imaging Make matter settings, the micro image acquisition control step, the imaging conditions be in the standby position is set samples were arranged in the image acquiring position for the micro image acquiring step, to acquire the micro image with reference to the image pickup condition In the semi-automatic mode, the imaging condition setting step displays the imaging condition automatically obtained corresponding to the macro image on the display means, and uses the imaging condition or manually sets the imaging condition. It is characterized by having the operator select whether to redo .

The image acquisition program according to the present invention includes (1) a sample storage unit for storing a plurality of samples, (2) a macro image acquisition unit for acquiring a macro image of the sample, and (3) a sample from the macro image. Is applied to an image acquisition apparatus including a micro image acquisition means for acquiring a micro image by scanning at a high resolution, and (4) a macro image acquisition control for controlling a macro image acquisition operation by the macro image acquisition means process, reference imaging condition setting processing for setting an image pickup condition for the micro image by the macro image, and includes a control processing with a micro image acquisition control processing for controlling an operation to acquire the micro image by the micro image acquiring means, image pickup condition setting The process refers to the macro image, sets the image acquisition range and the focus measurement position as the imaging conditions of the micro image, and the micro image acquisition control process Acquires focus information for the sample on the basis of the position, performs control to acquire the micro image based on the focus information and the image acquiring range obtained, (5) the control processing has a semi-automatic mode serving as a control mode (6) In the semi-automatic mode, the macro image acquisition control process acquires a macro image by arranging the sample at the storage position by the sample storage means at the image acquisition position by the macro image acquisition means for each of the plurality of samples. In addition, the imaging condition setting process is configured to allow the operator to confirm the macro image and the corresponding imaging condition for the sample from which the macro image has been acquired. sets a, after the macro image of the first sample has been obtained, in parallel with the acquisition of the macro image, to set the imaging conditions, Mi B image acquisition control processing performs control to an imaging condition In the standby position is set samples were placed at an image acquiring position for the micro image acquiring means, to acquire the micro image with reference to the image pickup condition, the semi-automatic In the mode, the imaging condition setting process displays the imaging condition automatically obtained corresponding to the macro image on the display means, and the operator determines whether to use the imaging condition or to manually reset the imaging condition. The computer is caused to execute a control process to be selected .

  In the image acquisition apparatus, the image acquisition method, and the image acquisition program described above, a macro image showing a whole image of the sample is provided by providing a macro image acquisition unit and a micro image acquisition unit for a plurality of samples to be acquired. The high-resolution micro image is acquired after setting the imaging conditions with reference to FIG. Thereby, for example, a high-resolution micro image of a sample that becomes image data of the sample used in a virtual microscope can be suitably acquired.

  In addition, as a control mode for controlling the acquisition of macro images, the setting of imaging conditions, and the acquisition of micro images for each sample, the operator must confirm the settings necessary to set the imaging conditions for micro images with reference to macro images. Semi-automatic mode to perform is provided. As a result, for example, even if there is an extra object such as dust in the sample slide, it is possible to reliably exclude the influence of such an object and correctly set the imaging conditions for the sample. .

  In the semi-automatic mode described above, the sample from which the macro image has been acquired by the macro image acquisition unit is not moved directly to the image acquisition position by the micro image acquisition unit, but is placed at the standby position. In such a configuration, acquisition of a macro image of a sample and setting of imaging conditions using the macro image can be performed independently of acquisition of a micro image that takes a certain amount of time to acquire a high-resolution image. It becomes. Thereby, after setting the imaging conditions for the sample, the operator does not need to be attached to the image acquisition device until the acquisition of the micro image is completed, thereby reducing the work load on the operator.

  In addition, about the stand-by position of the sample after macro image acquisition, it is good also as a structure which makes the storage position in a sample storage means the standby position, and returns the sample from which the macro image was acquired to a storage position again. Or it is good also as a structure which provides the sample waiting means which can be made to wait for a some sample separately from a sample storage means.

  In addition, the image acquisition apparatus includes a sample moving unit that moves each of the plurality of samples between the storage position, the standby position, and the image acquisition position in each of the macro image acquisition unit and the micro image acquisition unit. Is preferred. Similarly, the image acquisition method includes a sample moving step of moving each of the plurality of samples between a storage position, a standby position, and an image acquisition position in each of the macro image acquisition step and the micro image acquisition step. It is preferable. Similarly, the image acquisition program executes a sample movement process for moving each of the plurality of samples between the storage position, the standby position, and the image acquisition position in each of the macro image acquisition unit and the micro image acquisition unit. It is preferable to make it execute. Thereby, the acquisition of the macro image performed while moving the sample between the respective positions, the setting of the imaging conditions, and the acquisition of the micro image can be suitably realized.

  In addition, the image acquisition device includes an imaging device in which the micro image acquisition unit can acquire a one-dimensional image, or can acquire a two-dimensional image and perform TDI driving, and is orthogonal to the longitudinal direction of the imaging surface of the imaging device. The micro image acquisition control means scans the sample in the scanning direction with the imaging device to acquire a partial image, and the acquisition of the partial image is performed along the length of the imaging surface. It is preferable to perform control to acquire a plurality of partial images to be micro images by repeating a plurality of times while shifting the imaging position along the direction.

  Similarly, in the image acquisition method, the micro image acquisition step uses an imaging device capable of acquiring a one-dimensional image or capable of acquiring a two-dimensional image and TDI driving, and in the longitudinal direction of the imaging surface of the imaging device. The orthogonal direction is configured as the scanning direction when acquiring the micro image, and the micro image acquisition control step acquires the partial image by scanning the sample in the scanning direction by the imaging device, and acquires the partial image on the imaging surface. It is preferable to perform control to acquire a plurality of partial images to be micro images by repeating a plurality of times while shifting the imaging position along the longitudinal direction.

  Similarly, in the image acquisition program, the micro image acquisition unit has an imaging device capable of acquiring a one-dimensional image or capable of acquiring a two-dimensional image and TDI driving, and in the longitudinal direction of the imaging surface of the imaging device. The orthogonal direction is configured as a scanning direction when acquiring a micro image, and the micro image acquisition control process acquires a partial image by scanning the sample in the scanning direction by the imaging device, and acquires the partial image on the imaging surface. It is preferable to perform control to acquire a plurality of partial images to be micro images by repeating a plurality of times while shifting the imaging position along the longitudinal direction.

  According to such a configuration, a strip-shaped partial image obtained by scanning the sample in one direction with a one-dimensional sensor or a TDI drive two-dimensional sensor is obtained with high resolution, and a plurality of partial images are synthesized in the other direction to obtain a sample. By using the micro image of the entire image, it is possible to suitably acquire the image data of the sample with a sufficiently high resolution.

  Further, in the semi-automatic mode, the image acquisition device displays the imaging condition automatically obtained corresponding to the macro image on the display unit in the semi-automatic mode, and uses the imaging condition or manually sets the imaging condition. It is preferable to let the operator select whether to set again. Similarly, in the image acquisition method, in the semi-automatic mode, the imaging condition setting step displays the imaging condition automatically obtained corresponding to the macro image on the display unit, and uses the imaging condition or manually captures the image. It is preferable to let the operator select whether to reset the conditions. Similarly, in the semi-automatic mode, the image acquisition program causes the imaging condition setting process to display the imaging condition automatically obtained corresponding to the macro image on the display unit and use the imaging condition or manually capture the image. It is preferable to let the operator select whether to reset the conditions.

  According to such a configuration, for example, when there is a problem such as an extra object in the sample, the operator manually corrects or newly sets the imaging condition, and in other cases, the setting is automatically performed. It is possible to use a method in which the captured imaging conditions are used as they are. In this way, by manually setting the imaging conditions only when necessary, it is possible to efficiently perform image acquisition operations for a plurality of samples including the setting of the imaging conditions.

  Further, in the image acquisition apparatus, the control means has a fully automatic mode as its control mode, and in the fully automatic mode, the macro image acquisition control means takes the sample at the storage position as the image acquisition position in the macro image acquisition means. The imaging condition setting means automatically sets the imaging condition of the micro image corresponding to the macro image for the sample from which the macro image is acquired, and the micro image acquisition control means Alternatively, it is possible to perform control for arranging a sample for which imaging conditions are set at an image acquisition position in the micro image acquisition unit and acquiring a micro image with reference to the imaging conditions.

  Similarly, in the image acquisition method, the control step has a fully automatic mode as its control mode, and in the fully automatic mode, the macro image acquisition control step acquires the sample at the storage position in the macro image acquisition step. The imaging condition setting step automatically sets the imaging condition of the micro image corresponding to the macro image for the sample from which the macro image has been acquired, and performs the control for acquiring the macro image by arranging at the position. May be configured to place a sample for which an imaging condition is set at an image acquisition position in the micro image acquisition step, and to perform a control for acquiring a micro image with reference to the imaging condition.

  Similarly, in the image acquisition program, the control process has a fully automatic mode as its control mode. In the fully automatic mode, the macro image acquisition control process acquires an image of the sample at the storage position by the macro image acquisition means. The macro image acquisition control processing is performed by automatically setting the micro image imaging conditions corresponding to the macro image for the sample from which the macro image is acquired. May be arranged such that a sample for which an imaging condition is set is arranged at an image acquisition position in the micro image acquisition unit, and the computer performs control processing for performing control for acquiring a micro image with reference to the imaging condition.

  In such a configuration, as a control mode for controlling macro image acquisition, imaging condition setting, and micro image acquisition for each sample, either semi-automatic mode image acquisition or full-automatic mode image acquisition is operated. When the person appropriately selects, it is possible to execute image acquisition by a suitable method in accordance with the state of a plurality of samples to be acquired.

  In the image acquisition device, the imaging condition setting unit refers to the macro image and sets the image acquisition range and the focus measurement position as the imaging condition of the micro image, and the micro image acquisition control unit is based on the focus measurement position. It is preferable to perform control to acquire focus information for the sample and acquire a micro image based on the acquired focus information and an image acquisition range.

  Similarly, in the image acquisition method, the imaging condition setting step refers to the macro image, sets the image acquisition range and the focus measurement position as the micro image imaging condition, and the micro image acquisition control step is based on the focus measurement position. It is preferable to perform control for acquiring the focus information for the sample and acquiring the micro image based on the obtained focus information and the image acquisition range.

  Similarly, in the image acquisition program, the imaging condition setting process refers to the macro image, sets the image acquisition range and the focus measurement position as the imaging condition of the micro image, and the micro image acquisition control process is based on the focus measurement position. It is preferable to perform control for acquiring the focus information for the sample and acquiring the micro image based on the obtained focus information and the image acquisition range.

  Thereby, it is possible to suitably set parameters used for micro image acquisition from information about the sample obtained by the macro image, and to acquire image data of the sample in a good state at a high resolution.

  Further, regarding the configuration of the image acquisition means in the image acquisition device, the macro image acquisition means and the micro image acquisition means are a single image acquisition means, and the optical for image acquisition according to the acquisition of the macro image or the micro image The system may be configured to be switchable. An example of the switchable image acquisition optical system in this case is an optical system including a revolver including two types of objective lenses.

  According to the image acquisition device, the image acquisition method, and the image acquisition program of the present invention, the macro image acquisition, the imaging condition setting, and the micro image acquisition control for each of the plurality of samples are referred to the macro image. In addition to providing a semi-automatic mode for the operator to perform necessary confirmation when setting the imaging conditions for micro images, without moving the sample from which the macro image was acquired directly to the image acquisition position in the micro image acquisition means, By arranging at the standby position, it is possible to efficiently execute image acquisition for each of the plurality of samples.

  Hereinafter, preferred embodiments of an image acquisition device, an image acquisition method, and an image acquisition program according to the present invention will be described in detail with reference to the drawings. In the description of the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted. Further, the dimensional ratios in the drawings do not necessarily match those described.

  First, the overall configuration of the image acquisition device will be described. FIG. 1 is a block diagram showing a configuration of an embodiment of an image acquisition apparatus according to the present invention. The image acquisition apparatus according to the present embodiment is a microscope system used for acquiring an image of the sample S with high resolution. The microscope unit 10 acquires an image of the sample S, and controls image acquisition in the microscope unit 10. The control part 60 to perform is provided. An example of the sample S that is an object of image acquisition is a slide (preparation) in which a biological sample such as a tissue section is sealed on a slide glass when acquiring image data used in a virtual microscope.

  The microscope unit 10 includes a sample storage unit 11, a macro image acquisition unit 20, and a micro image acquisition unit 30. The sample storage unit 11 is storage means configured to be capable of storing a plurality of samples (for example, a plurality of slides each sealed with a biological sample) S that are objects of image acquisition. The sample storage unit 11 is provided with a door 12 used for storing and taking out the sample S by an operator. In the present embodiment, an interlock mechanism 13 is provided to prevent the door 12 from being accidentally opened during image acquisition.

  The macro image acquisition unit 20 is a first image acquisition unit for acquiring a macro image as a low-magnification image of the sample S. In the image acquisition unit 20, a macro image with a low resolution corresponding to the entire image of the sample S is acquired. Further, a macro light source 25 that supplies light for generating a light image of the sample S at the time of macro image acquisition is installed in the macro image acquisition unit 20.

  On the other hand, the micro image acquisition unit 30 is a second image acquisition unit for acquiring a micro image as a high-magnification image of the sample S. In the image acquisition unit 30, a micro image of the target sample S at a high resolution is acquired. Further, a micro light source 35 that supplies light for generating a light image of the sample S at the time of micro image acquisition is installed in the micro image acquisition unit 30. Further, in FIG. 1, an image correction unit 38 for performing correction necessary for the image data is provided for the micro image acquired by the image acquisition unit 30. Examples of correction performed by the image correction unit 38 include dark correction and shading correction. For example, when performing shading correction, a method can be used in which a blank image obtained by imaging a reference sample such as a slide on which a biological sample is not placed is acquired in advance, and shading correction is performed with reference to the blank image. . These image corrections may be performed in the same manner for the macro image.

  A sample transport unit 14 and a sample stage 15 are provided as sample moving means for moving the sample S between positions in the microscope unit 10. The sample transport unit 14 is a transport unit that transports the sample S between a storage position in the sample storage unit 11 and an image acquisition position in each of the macro image acquisition unit 20 and the micro image acquisition unit 30. The sample stage 15 is used for setting and adjusting the image acquisition position of the sample S while the sample S is placed when acquiring a macro image or a micro image. Further, the microscope unit 10 is provided with a drive control unit 40 for driving and controlling each unit of the microscope unit 10.

  The control unit 60 includes a data processing unit 70, a data storage unit 75, and an image acquisition control unit 80. The data processing unit 70 is input with image data of the macro image acquired by the macro image acquisition unit 20 and image data of the micro image acquired by the micro image acquisition unit 30, and necessary for these image data. Data processing is performed.

  The image acquisition control unit 80 controls the image acquisition operation of the sample S in the microscope unit 10 via the drive control unit 40. Further, image data input to the data processing unit 70, various data and information obtained by processing the image data, control information used in the image acquisition control unit 80, and the like are stored in the data storage unit 75 as necessary. Is stored and retained.

  The control unit 60 is configured by a computer including a CPU and a storage device such as a necessary memory and a hard disk. A display device 61 and an input device 62 are connected to the control unit 60. The display device 61 is, for example, a CRT display or a liquid crystal display, and is used to display an operation screen necessary for the operation of the image acquisition device or an image of the acquired sample S. The input device 62 is, for example, a keyboard or a mouse, and is used for inputting information necessary for image acquisition, inputting instructions for an image acquisition operation, and the like.

  Next, the configuration of the microscope unit 10 in the image acquisition apparatus illustrated in FIG. 1 will be described. FIG. 2 is a diagram schematically showing the configuration of the microscope unit 10. As shown in FIG. 2, the microscope unit 10 according to the present embodiment is configured as a transmission type microscope system used for acquiring a light image of the sample S. Here, as shown in the figure, two directions orthogonal to each other in the horizontal direction are defined as an X-axis direction and a Y-axis direction, and a vertical direction orthogonal to the horizontal direction is defined as a Z-axis direction. Of these, the Z-axis direction, which is the vertical direction, is the direction of the optical axis for image acquisition in the microscope system. 2 mainly shows the configuration of the macro image acquisition unit 20 and the micro image acquisition unit 30, and the sample storage unit 11, the sample transport unit 14, and the like are not shown.

  The sample S is placed on the sample stage 15 when the image acquisition units 20 and 30 acquire images. The sample stage 15 is configured as an XY stage that can move in the X-axis direction and the Y-axis direction using a stepping motor, a DC motor, a servo motor, or the like. In such a configuration, the image acquisition position with respect to the sample S in the image acquisition units 20 and 30 is set and adjusted by driving the sample stage 15 in the XY plane. In the present embodiment, the sample stage 15 is movable between an image acquisition position in the macro image acquisition unit 20 and an image acquisition position in the micro image acquisition unit 30.

  The macro image acquisition unit 20 and the macro light source 25 are installed at predetermined positions on the optical axis 20a with respect to the macro image acquisition position for acquiring the macro image of the sample S, respectively. The macro light source 25 is a light source that irradiates the sample S with light for generating a macro image acquisition light image, and is installed below the sample stage 15.

  The macro image acquisition unit 20 is configured by using an imaging device 21 such as a two-dimensional CCD sensor that can acquire a two-dimensional image from an optical image of the sample S. An imaging optical system 22 is provided between the macro image acquisition position where the sample S is arranged and the imaging device 21 as an optical system for guiding the optical image of the sample S.

  On the other hand, with respect to the micro image acquisition position for acquiring the micro image of the sample S, the micro image acquisition unit 30 and the micro light source 35 are installed at predetermined positions on the optical axis 30a. The micro light source 35 is a light source for irradiating the sample S with light for generating a light image for acquiring a micro image, and is installed below the sample stage 15 together with the condenser lens 36.

  The micro image acquisition unit 30 is configured by using an imaging device 31 such as a one-dimensional CCD sensor that can acquire a one-dimensional image from an optical image of the sample S. Further, an objective lens 32 and a light guide optical system 34 are provided as an optical system for guiding an optical image of the sample S between the micro image acquisition position where the sample S is arranged and the imaging device 31. The objective lens 32 receives the light transmitted through the sample S and generates an optical image of the sample S. Further, the light guide optical system 34 is constituted by, for example, a tube lens, and guides the optical image of the sample S to the imaging device 31.

  Further, a Z stage 33 using a stepping motor or a piezo actuator is provided for the objective lens 32, and the objective lens 32 is driven in the Z-axis direction by the Z stage 33, thereby focusing on the sample S. Etc. are possible. The imaging device 31 in the micro image acquisition unit 30 is not limited to an imaging device capable of acquiring a one-dimensional image, and an imaging device capable of acquiring a two-dimensional image and TDI driving can also be used. .

  Further, as the imaging device 31 for acquiring a micro image, it is preferable to use an imaging device capable of acquiring a color image such as a three-plate CCD camera. Note that as the imaging device 21 for acquiring a macro image, either an imaging device for acquiring a monochrome image or an imaging device capable of acquiring a color image may be used as necessary. In addition, as for the imaging device 31, when it is not necessary to acquire a color image, an imaging device for acquiring a monochrome image may be used.

  In contrast to the sample stage 15, the macro image acquisition unit 20, the micro image acquisition unit 30, and the light sources 25 and 35, the drive control unit 40 includes a stage control unit 41, an imaging device control unit 42, and a light source control unit 43. Is provided. The stage control unit 41 sets and adjusts the imaging condition for the sample S by driving and controlling the sample stage 15 and the Z stage 33 which are XY stages. Further, the imaging device control unit 42 controls the image acquisition of the sample S by controlling the driving of the imaging devices 21 and 31. The light source control unit 43 controls the irradiation of the image acquisition light to the sample S by driving and controlling the light sources 25 and 35. In addition, about control of each part of the microscope part 10, you may use other structures, such as the structure directly controlled by the image acquisition control part 80 of the control part 60 without going through the drive control part 40.

  Here, acquisition of the macro image and the micro image of the sample S in the image acquisition units 20 and 30 will be described. The macro image acquisition unit 20 acquires a macro image that is an overall image of the sample S used for setting the imaging conditions of the micro image. For example, when a slide in which a biological sample or the like is sealed on the above-described slide glass is used as the sample S, the entire slide or an image in a predetermined range including the biological sample is acquired as the macro image.

  The micro image acquisition unit 30 acquires a micro image of the sample S at a target resolution with reference to the set imaging conditions. The acquisition of the micro image is performed by two-dimensionally scanning the sample S at a predetermined resolution higher than that of the macro image, as schematically shown in FIG. Here, in the acquisition of a micro image using the imaging device 31 such as a one-dimensional CCD camera, the longitudinal direction of the imaging surface of the imaging device 31 in the XY plane parallel to the sample S is the X-axis direction. A direction orthogonal to the direction is defined as a Y-axis direction. At this time, in the micro image acquisition, the direction perpendicular to the longitudinal direction of the imaging surface of the imaging device 31 and the negative direction of the Y axis in FIG.

  In acquiring a micro image using an imaging device 31 such as a one-dimensional CCD camera, first, the sample S on the sample stage 15 is scanned in the scanning direction (negative direction of the Y axis) by the imaging device 31 to obtain a desired resolution. A strip-shaped partial image A having Further, as shown in FIG. 3A, the acquisition of such partial images is repeated a plurality of times while shifting the imaging position along the longitudinal direction of the imaging surface (the positive direction of the X-axis) to obtain a plurality of partial images. A, B, ..., I are acquired.

  By combining the partial images A to I obtained in this way in the X-axis direction, a micro image of the entire sample S can be generated. According to such a micro image acquisition method, the image data of the sample S can be preferably acquired with sufficiently high resolution. In FIG. 3A, an area having the X-axis direction indicated by hatching in the partial image A as the longitudinal direction indicates an imaging area corresponding to the imaging surface of the imaging device 31.

  In addition, regarding the setting of the micro image capturing condition, the image acquisition range and the focus measurement position are set as the micro image capturing condition with reference to the macro image acquired by the imaging device 21 of the macro image acquiring unit 20. Is preferred. As a result, it is possible to suitably set the parameters used for micro image acquisition from the information obtained in the macro image that is the entire image of the sample S, and to acquire the image data of the sample in a good state with high resolution. Become.

  Specifically, when the slide is the sample S as described above, as shown in FIG. 3B, the image acquisition range for the sample S is a rectangle including the biological sample L in the slide that is the target of image acquisition. It can be set by the shape range R. Two-dimensional scanning (see FIG. 3A) of the sample S in the micro image acquisition unit 30 is performed within the image acquisition range R set in this way. When such an image acquisition range R is automatically set, for example, an image acquisition target is obtained by binarizing an image with reference to a threshold value set for a luminance pattern in a macro image. A method of determining a range where an object (for example, a biological sample L) exists and setting the image acquisition range R based on the determination result can be used.

  FIG. 4 is a graph showing a threshold setting method for binarizing a macro image. In the graph of FIG. 4, the horizontal axis indicates the luminance value at each pixel of the macro image. Here, in the macro image acquired in the present embodiment, due to the transmission type configuration of the macro image acquisition unit 20, the luminance is highest in the region of only the slide glass where the biological sample L does not exist.

  In setting the threshold value for the macro image, for example, as shown in FIG. 4, a method of obtaining two peaks P1 and P2 in the luminance distribution and setting the luminance value at the center as the threshold value T can be used. Then, by binarizing the image based on this threshold, it is possible to easily determine the range where the object L exists in the image. Further, among the determined ranges, a continuous range that is equal to or larger than the specified area is set as the range of one object L, and the smallest rectangular area that includes all the objects L is set as the image acquisition range R. In addition to the method described above, various methods can be used for setting the threshold, for example, a method of setting the threshold at a fixed ratio with respect to the peak luminance value.

  Further, in the determination of the range in which the object L exists in the binarized macro image, n times of contraction processing (erosion processing: processing for reducing a picture block) is performed on the binarized image. Subsequently, it is preferable to perform n times of expansion processing (dilation processing: processing for enlarging a block of pictures). Thereby, small noises and the like in the image can be removed. Further, a filtering process for removing the edge of the cover glass of the slide or dust in the slide may be further performed.

  The focus measurement position is used when the micro image acquisition unit 30 acquires focus information for the sample S prior to acquisition of the micro image of the sample S. The micro image acquisition unit 30 performs focus measurement on the set focus measurement position using the imaging device 31 to determine a focus position as focus information when acquiring a micro image of the sample S. For example, when the tilt of the sample S in the horizontal plane, that is, the shift of the focus position in the horizontal plane does not matter, the focus measurement position may be set to one focus measurement position for the sample S.

  Further, when it is necessary to consider the shift of the focus position in the horizontal plane, it is preferable to set three or more focus measurement positions for the sample S. Thus, by performing focus measurement by setting three or more focus measurement positions, a two-dimensional focus map for the image acquisition range R of the sample S can be obtained. For example, when the focal map for the focal position is determined as a planar focal plane, the focal plane can be obtained from the plane including the measurement result points at the three focal measurement positions. When four or more focus measurement positions are used, the focal plane can be obtained from the measurement result points using a fitting method such as a least square method.

  FIG. 3B shows an example in which nine focus measurement positions are automatically set for setting a focus measurement position using a macro image. Here, the image acquisition range R previously set for the sample S is divided into 9 equal parts by 3 × 3, and nine focus measurement positions P are set by the center points of the respective areas.

  In addition, here, among the nine focus measurement positions, eight of the initial positions are included in the range of the biological sample L that is the object of image acquisition, and thus are set as the focus measurement positions as they are. The On the other hand, the lower left point is outside the range of the biological sample L, and cannot be used as a focus measurement position as it is. For this reason, for the focus measurement position at the lower left, for example, a position Q obtained by a method such as moving toward the center within the image acquisition range R may be set as the focus measurement position. Alternatively, such a position may be excluded from the focus measurement position.

  In addition, when obtaining a focal plane using the least square method from four or more focus measurement positions, if there is a measurement position that is too far from the focal plane obtained in the focus measurement position, except that, again, It is preferable to re-determine the focal plane. Further, when the focal plane cannot be normally obtained, it is preferable to exclude the object as dust.

  As in the example described above, when the sample S is a slide, with respect to the imaging conditions for acquiring the micro image, preferably, referring to the macro image acquired by the macro image acquisition unit 20 first, the micro image is obtained. As an image capturing condition, an image acquisition range R including the biological sample L and a focus measurement position P with a predetermined number of points are set. Then, the micro image acquisition unit 30 acquires focus information about a focus position or a focal plane with respect to the sample S based on the focus measurement position P, and based on the obtained focus information and the set image acquisition range R. The micro image of the sample S is acquired.

  The setting of the image acquisition range R and the focus measurement position P using the macro image of the sample S is not limited to the example illustrated in FIG. 3B, and various methods may be used specifically. For example, FIG. 3B shows an example in which the focus measurement position P is automatically set using a predetermined setting algorithm. However, when manually setting the focus measurement position, the operator must After confirming the macro image, the focus measurement position may be set so as to obtain an appropriate number of points and arrangement.

  Next, the configuration of the control unit 60 in the image acquisition apparatus illustrated in FIG. 1 will be described. FIG. 5 is a block diagram illustrating a configuration of the control unit 60. As shown in FIG. 5, the control unit 60 according to the present embodiment includes a data processing unit 70, a data storage unit 75, and an image acquisition control unit 80.

  The data processing unit 70 includes an imaging condition setting unit 71 and a sample data creation unit 72. The imaging condition setting unit 71 is a setting unit that sets the imaging condition of the micro image with reference to the macro image acquired by the macro image acquisition unit 20 of the microscope unit 10. The sample data creating unit 72 creates sample data that is image data of the sample S using the micro image acquired by the micro image acquiring unit 30.

  FIG. 6 is a diagram schematically illustrating creation of sample data using a micro image. Here, the image data group of strip-shaped partial images A, B, C,... Is input to the control unit 60 as the image data of the micro image acquired by the micro image acquisition unit 30 of the microscope unit 10 (FIG. 3). (See (a)). The sample data creation unit 72 synthesizes these partial images side by side, generates image data that becomes a micro image of the entire sample S, and uses it as sample data. This sample data can be used as image data in a virtual microscope, for example. Note that the image data of the sample S may be compressed as necessary. The image data input from the microscope unit 10 and the sample data created by the sample data creation unit 72 are stored in the data storage unit 75 as necessary.

  The image acquisition control unit 80 includes a macro image acquisition control unit 81 and a micro image acquisition control unit 82. The macro image acquisition control unit 81 controls the macro image acquisition operation of the sample S by the macro image acquisition unit 20. Further, the micro image acquisition control unit 82 controls the micro image acquisition operation by the micro image acquisition unit 30. The control of the acquisition operation of the macro image and the micro image of the sample S by the image acquisition control units 81 and 82 and the setting of the imaging condition by the imaging condition setting unit 71 are performed according to the control mode selected for the image acquisition process. . In the present embodiment, the control unit 60 has three control modes, a fully automatic mode, a manual mode, and a semi-automatic mode, as its control mode.

  FIG. 7 is a flowchart showing an image acquisition method in the fully automatic mode. In the fully automatic mode, the macro image acquisition control unit 81 of the control unit 60 controls to acquire the macro image by arranging the sample S in the storage position in the sample storage unit 11 at the image acquisition position in the macro image acquisition unit 20. (Macro image acquisition control step). Further, the imaging condition setting unit 71 automatically sets the imaging condition of the micro image corresponding to the macro image for the sample S from which the macro image is acquired (imaging condition setting step). Further, the micro image acquisition control unit 82 performs control to place the sample S for which the imaging condition is set at the image acquisition position in the micro image acquisition unit 30 and acquire the micro image with reference to the imaging condition (micro image). Acquisition control step).

  In the example shown in FIG. 7, first, the operator sets a plurality of slides S to be imaged in the sample storage unit 11 of the microscope unit 10 and closes the door 12 (step S101). The number of slides that can be stored in the sample storage unit 11 varies depending on the specific apparatus configuration, but is about several hundreds, for example.

  Next, among the plurality of slides S, the slide S for performing image acquisition is taken out from the sample storage unit 11, transported by the sample transport unit 14, and placed on the sample stage 15. The image acquisition position (see FIG. 1 and FIG. 2) is set (S102). Then, a macro image of the slide S including the biological sample L is acquired by the imaging device 21 of the image acquisition unit 20 (S103, macro image acquisition step). The imaging condition setting unit 71 of the control unit 60 refers to the acquired macro image, and an image acquisition range R that is an imaging condition for the slide S and a plurality of focus measurement positions P are automatically determined using a predetermined algorithm. (S104, imaging condition setting step).

  The slide S for which the acquisition of the macro image is completed is moved from the image acquisition position in the macro image acquisition unit 20 by the sample transport unit 14 or the sample stage 15 and is set to the image acquisition position in the micro image acquisition unit 30 ( S105). Then, with reference to the imaging conditions set by the imaging condition setting unit 71, the micro image of the slide S is acquired by the imaging device 31 of the image acquisition unit 30 (S106, micro image acquisition step).

  Specifically, first, the micro image acquisition unit 30 performs focus measurement for each of the plurality of focus measurement positions P set for the slide S, and is optimal for acquiring an image of the biological sample L from the obtained focus position. The focal plane is obtained as the correct focus information. Then, while performing focus control based on the determined focal plane, the image acquisition range 31 scans the slide S two-dimensionally with the imaging device 31 to acquire a plurality of strip-shaped partial images. The plurality of partial images become high-resolution micro images of the slide S (digital slides in a virtual microscope) by performing predetermined data synthesis processing in the sample data creation unit 72 of the control unit 60.

  Subsequently, the slide S for which micro image acquisition has been completed is returned from the image acquisition position in the micro image acquisition unit 30 to the storage position in the sample storage unit 11 by the sample transport unit 14 (S107). Then, the image acquisition control unit 80 checks whether or not there is a slide S for which image acquisition processing has not been completed among the plurality of slides S (S108). Here, if there is a slide S to be subjected to further image acquisition processing, the above steps S102 to S107 are repeated. If the image acquisition processing for all the slides S has been completed, the image acquisition for the plurality of slides S set in the sample storage unit 11 is terminated.

  The manual mode is basically the same as the image acquisition procedure in the fully automatic mode except that each step is manually performed according to an instruction from the operator. However, even in the manual mode, for example, steps that do not require an instruction from the operator, such as movement of the slide S for which the macro image has been acquired to the image acquisition position in the micro image acquisition unit 30, are automatically performed. Also good. Further, regarding the acquisition of the micro image of the slide S, the operator can confirm the acquired micro image, and if there is a problem, the imaging condition can be reset or the micro image can be re-acquired. There may be.

  8 and 9 are flowcharts illustrating an image acquisition method in the semi-automatic mode. In the semi-automatic mode, the macro image acquisition control unit 81 of the control unit 60 places the sample S at the storage position in the sample storage unit 11 at the image acquisition position in the macro image acquisition unit 20 for each of the plurality of samples S. Then, a macro image is acquired, and further control is performed to place the sample S at the standby position (macro image acquisition control step). Further, the imaging condition setting unit 71 sets the imaging condition of the micro image by allowing the operator to confirm the macro image and the imaging condition corresponding thereto for the sample S from which the macro image is acquired (imaging condition setting step). Further, the micro image acquisition control unit 82 arranges the sample S at the standby position and for which the imaging condition is set at the image acquisition position in the micro image acquisition unit 30, and acquires the micro image with reference to the imaging condition. (Micro image acquisition control step). In the present embodiment, the standby position of the sample S is set to the storage position in the sample storage unit 11.

  In the example shown in FIGS. 8 and 9, first, similarly to the case of the fully automatic mode, the operator sets a plurality of slides S to be imaged in the sample storage unit 11 of the microscope unit 10, and the door 12. Is closed (step S201).

  Next, the macro image acquisition control unit 81 acquires the macro image for each of the plurality of slides S (S202, macro image acquisition step). Specifically, as shown in FIG. 8, among the plurality of slides S, the slide S for executing image acquisition is taken out from the storage position in the sample storage unit 11, transported by the sample transport unit 14, and sample stage 15. The image is placed on the image acquisition position in the macro image acquisition unit 20 (S301). Then, the macro image of the slide S including the biological sample L is acquired by the imaging device 21 of the image acquisition unit 20 (S302).

  The slide S for which the macro image has been acquired is returned from the image acquisition position in the macro image acquisition unit 20 to the storage position in the sample storage unit 11 that is the standby position by the sample transport unit 14 (S303). Then, the macro image acquisition control unit 81 checks whether or not there is a slide S for which the macro image acquisition process has not been completed among the plurality of slides S (S304). If there is a slide S to be subjected to further macro image acquisition processing, the above steps S301 to S303 are repeated. If the macro image acquisition process for all the slides S has been completed, the macro image acquisition for the plurality of slides S set in the sample storage unit 11 is terminated.

  Further, in this mode, after the macro image of the first slide S is acquired, the imaging conditions for the micro image are set in parallel with the macro image acquisition process (S203, imaging condition setting step). The imaging condition setting unit 71 of the control unit 60 obtains a micro image by causing the display means to confirm the acquired macro image and the corresponding imaging condition for the slide S for which the imaging condition is to be set. An image acquisition range R and a plurality of focus measurement positions P, which are image capturing conditions, are set. The imaging condition setting method here will be described later in detail.

  When the macro image acquisition processing for all the slides S is completed, the micro image acquisition control unit 82 acquires the micro images for each of the plurality of slides S in parallel with the imaging condition setting processing. (S204, micro image acquisition step). Specifically, as shown in FIG. 9, among the plurality of slides S, a slide S for which imaging conditions have already been set is selected, and image acquisition is executed. The selected slide S is taken out from the storage position in the sample storage unit 11 which is the standby position, transported by the sample transport unit 14 and placed on the sample stage 15, and set at the image acquisition position in the micro image acquisition unit 30. (S401). Then, with reference to the imaging conditions set by the imaging condition setting unit 71, the micro image of the slide S is acquired by the imaging device 31 of the image acquisition unit 30 (S402). Note that the micro image acquisition method is basically the same as in the fully automatic mode.

  Subsequently, the slide S for which micro image acquisition has been completed is returned from the image acquisition position in the micro image acquisition unit 30 to the storage position in the sample storage unit 11 by the sample transport unit 14 (S403). Then, in the micro image acquisition control unit 82, it is confirmed whether or not there is another slide S for which the imaging condition is set among the plurality of slides S (S404). If there is a slide S for which image acquisition conditions are further set and image acquisition processing is possible, the above steps S401 to S403 are repeated.

  If there is no slide S for which the imaging condition is set, the micro image acquisition control unit 82 further confirms whether there is a slide S for which image acquisition processing has not been completed among the plurality of slides S. (S405). Here, if there is a slide S to be subjected to further image acquisition processing, the above-described steps S401 to S403 are repeatedly executed after the imaging condition is set. If the image acquisition processing for all the slides S has been completed, the image acquisition for the plurality of slides S set in the sample storage unit 11 is terminated.

  In the image acquisition apparatus of the present embodiment, a session is defined corresponding to a plurality of samples S set in the sample storage unit 11 by an operator, a sample group of the plurality of samples S, and a corresponding macro image. Data groups such as imaging conditions and micro images are associated by this session. Corresponding to such a configuration, the control unit 60 shown in FIG. 5 is provided with a session management unit 73 in the data processing unit 70 and a session switching unit 83 in the image acquisition control unit 80.

  The session management unit 73 manages data groups such as macro images, imaging conditions, and micro images for each session, and stores them in the data storage unit 75 as necessary. In the data storage unit 75 of FIG. 5, as an example of such data management, a case is shown in which data groups of a plurality of sessions of session 1, session 2,. In addition, the session switching unit 83 controls switching of image acquisition processing associated with exchange of sample groups of a plurality of samples S, and instructs the session management unit 73 to switch corresponding data groups. As described above, by associating the sample groups of the plurality of samples S with the data groups in the image acquisition using the concept of session, the control of the image acquisition processing of the samples S and the management of the data corresponding thereto are suitably performed. It can be carried out.

  A specific example of setting a micro-image imaging condition in the semi-automatic mode will be described. FIG. 10 is a flowchart illustrating an example of an imaging condition setting method. FIGS. 11 to 13 are diagrams illustrating examples of operation screens used for setting the imaging conditions. Note that these operation screens are displayed on the display device 61 in the configuration shown in FIG.

  In a state where a macro image is acquired for one or a plurality of slides S among a plurality of slides S to be acquired, an operation screen 100 shown in FIG. On the operation screen 100, in the list display unit 101, a list of slides from which macro images have been acquired (slides 1 to 3 in FIG. 11) is displayed together with thumbnail images of the corresponding macro images.

  In addition, the operator clicks a desired slide name, for example, “slide 1” on the screen 100, thereby expanding the macro image of the selected “slide 1” to the macro image display unit 102 as shown in FIG. You can display it and check its contents. Then, by clicking the imaging condition setting button 103 in this state, a slide for setting imaging conditions is selected from the displayed slide list (step S501).

  When a slide for setting imaging conditions is selected, a macro image of the selected slide 1 is displayed on the macro image display unit 111 in the imaging condition setting screen 110 shown in FIG. The operator confirms the macro image and selects whether to set the imaging condition automatically or manually (S502). Then, it is confirmed whether or not the automatic setting is selected (S503). If the automatic setting is selected, the imaging condition is automatically set and the setting of the imaging condition for the slide is ended, and the next slide is selected. , And the imaging condition setting operation.

  If the manual setting is selected, the operator sets the image acquisition range R and the focus measurement position P for the macro image of the slide in consideration of the range and shape of the biological sample L. Set manually (S504). When the setting of the imaging condition is completed, it is confirmed whether there is a slide S for which the setting of the imaging condition has not been completed among the plurality of slides S (S505). Here, if there is a slide S for which imaging conditions are further set, steps S501 to S504 described above are repeated. If the setting of the imaging conditions for all the slides S has been completed, the setting of the imaging conditions for the plurality of slides S set in the sample storage unit 11 is completed.

  In the example of the imaging condition setting screen 110 shown in FIG. 13, when the macro image of the slide S is displayed on the macro image display unit 111, the image acquisition range R and the focus measurement position P by automatic setting are first obtained. These are displayed on the macro image as the initial setting of the imaging conditions. The operator confirms the automatically set imaging condition displayed here, and selects whether to set the imaging condition automatically or manually. When the imaging condition is automatically set, the initial setting displayed first may be used as it is, so the setting end button 112 is clicked, and the imaging condition setting for the slide S is completed.

  Further, the automatically acquired image acquisition range R and focus measurement position P may be displayed on the thumbnail image of the macro image in the list display unit 101 shown in FIG. As described above, by superimposing the imaging conditions on the thumbnail image, the imaging conditions of each slide can be listed, so that a slide to be manually set can be easily extracted.

  When manually setting the imaging conditions, the focus measurement position setting button 113 or the image acquisition range setting button 114 is clicked as necessary. Then, the imaging conditions are manually set by performing operations such as changing, adding, deleting, or creating a new focus measurement position P or image acquisition range R on the macro image display unit 111. If necessary, the imaging condition can be automatically set again by clicking the automatic setting button 115. When the setting of the imaging condition is completed, the setting end button 112 is clicked, and the setting of the imaging condition for the slide S is completed.

  The process corresponding to the image acquisition method executed in the image acquisition apparatus shown in FIG. 1 can be realized by an image acquisition program for causing a computer to execute the image acquisition process. For example, the control unit 60 of the image acquisition device stores a CPU that operates each software program necessary for image acquisition processing, a ROM that stores the software program, and the like, and data is temporarily stored during execution of the program. It can be constituted by a RAM. In such a configuration, the above-described image acquisition device and image acquisition method can be realized by executing a predetermined image acquisition program by the CPU.

  Further, the above-described program for causing the CPU to execute each process for obtaining the image of the sample can be recorded on a computer-readable recording medium and distributed. In such a recording medium, for example, a magnetic medium such as a hard disk and a flexible disk, an optical medium such as a CD-ROM and a DVD-ROM, a magneto-optical medium such as a floppy disk, or a program instruction is executed or stored. For example, hardware devices such as RAM, ROM, and semiconductor non-volatile memory are included.

  The effects of the image acquisition device, the image acquisition method, and the image acquisition program according to the present embodiment will be described.

  In the image acquisition device, the image acquisition method, and the image acquisition program described above, the macro image acquisition unit 20 and the micro image acquisition unit 30 are provided for the plurality of samples S that are the target of image acquisition, and the imaging condition setting unit 71. In FIG. 5, a high-resolution micro image is acquired after setting imaging conditions with reference to a macro image showing the entire image of the sample S. Thereby, for example, a high-resolution micro image of a sample that becomes image data of a sample used in a virtual microscope can be suitably acquired.

  Further, the control unit 60 that controls the image acquisition operation in the microscope unit 10 refers to the macro image as a control mode for controlling the acquisition of the macro image, the setting of the imaging condition, and the acquisition of the micro image for each sample S. A semi-automatic mode is provided in which the operator performs necessary confirmation when setting the micro-image capturing conditions. As a result, for example, even if there is an extra object such as writing of information about the biological sample or dust in the slide as the sample S, the influence of such an object in the setting of the imaging condition is surely excluded. Thus, it is possible to correctly set the imaging conditions for the sample.

  Further, in the semi-automatic mode described above, the sample S from which the macro image is acquired by the macro image acquisition unit 20 is not directly moved to the image acquisition position by the micro image acquisition unit 30 but is arranged at the standby position. . In such a configuration, as shown in the flowcharts of FIGS. 8 and 9, the acquisition of the macro image of the sample S (step S202) and the setting of the imaging condition using the macro image (S203) Can be performed independently of micro-image acquisition (S204), which takes a certain amount of time to scan the sample S.

  Thereby, the operator refers to the set imaging condition for obtaining the micro image after completing the acquisition of the macro image for each of the plurality of samples S and the setting and confirmation of the imaging condition using the macro image. Since it is automatically executed by the micro image acquisition control unit 82, it is not necessary to be attached to the image acquisition device until the acquisition of the micro image is completed. As a result, the burden on the operator is greatly reduced.

  As for the standby position of the sample S after obtaining the macro image, in the above embodiment, the storage position in the sample storage unit 11 is set as the standby position, and the sample S from which the macro image is acquired is returned to the storage position again. Yes. Thereby, the structure of the microscope part 10 can be simplified. Or it is good also as a structure which provides the sample stand-by part which can make the some sample S stand by separately from the sample storage part 11. FIG.

  Moreover, the above-described image acquisition apparatus includes a plurality of samples S, each of which stores a storage position in the sample storage unit 11, a standby position (in the above configuration, matches the storage position), a macro image acquisition unit 20, and a micro image acquisition unit. As a sample moving means for moving between 30 image acquisition positions, a sample transport unit 14 and a sample stage 15 that is an XY stage are provided. With such a configuration, it is possible to suitably realize macro image acquisition, imaging condition setting, and micro image acquisition performed while moving the sample S between the positions.

  The micro image acquisition unit 30 includes an imaging device 31 including a one-dimensional sensor capable of acquiring a one-dimensional image or a TDI-driven two-dimensional sensor capable of acquiring a two-dimensional image and TDI driving. The direction perpendicular to the longitudinal direction of the imaging surface at is configured as a scanning direction when acquiring a micro image. Furthermore, the micro image acquisition control unit 82 acquires a partial image by scanning the sample S in the scanning direction by the imaging device 31, and acquires the partial image a plurality of times while shifting the imaging position along the longitudinal direction of the imaging surface. Repeatedly, control is performed to acquire a plurality of partial images to be micro images. According to such a configuration, a strip-shaped partial image obtained by scanning the sample S in one direction is acquired with high resolution, and a plurality of partial images are synthesized in the other direction to form a micro image of the entire sample. The image data of the sample S can be suitably acquired with sufficiently high resolution.

  In the image acquisition device described above, in the semi-automatic mode, the imaging condition setting unit 71 displays the imaging condition automatically obtained corresponding to the macro image on the display device 61 as the initial setting of the imaging condition. The operator is allowed to select whether to use the imaging conditions or to manually reset the imaging conditions. According to such a configuration, for example, when there is a problem such as an extra object on the sample S, the operator manually corrects or newly sets the imaging condition, and in other cases, the imaging condition is automatically set. A method of using the set imaging conditions as they are can be used. As described above, by manually setting the imaging conditions only when necessary, it is possible to efficiently perform the image acquisition operation of the plurality of samples S including the setting of the imaging conditions.

  In the image acquisition device described above, the imaging condition setting unit 71 refers to the macro image, sets the image acquisition range R and the focus measurement position P as the imaging conditions for the micro image, and the micro image acquisition control unit 82 The focus information for the sample S is acquired based on the focus measurement position P, and control for acquiring a micro image based on the obtained focus information and the image acquisition range R is performed. Accordingly, it is possible to appropriately set parameters used for micro image acquisition from information about the sample S obtained as a macro image, and to acquire image data of a sample in a good state with high resolution.

  Further, as a control mode of the image acquisition operation by the control unit 60, a fully automatic mode is prepared in addition to the semi-automatic mode. In such a configuration, as a control mode for controlling macro image acquisition, imaging condition setting, and micro image acquisition for each sample, either semi-automatic mode image acquisition or full-automatic mode image acquisition is operated. By appropriately selecting an image, it is possible to perform image acquisition by a suitable method according to the state of the plurality of samples S to be acquired, for example, whether or not each sample S has a lot of dust. Become.

  Note that these control modes may be configured such that the control modes can be switched, such as switching to the fully automatic mode during execution of the semi-automatic mode. In both the fully automatic mode and the semi-automatic mode, if there is a problem with the acquired micro image after completing the image acquisition process, the image acquisition for the sample S can be performed again. Is preferred.

  Also, when exactly the same conditions can be set for a plurality of slides, a condition is set manually for the first slide, and the set conditions are applied to the remaining slides. It is also good. Alternatively, the conditions set in advance in the control unit 60 may be applied to one or a plurality of slides. Moreover, after performing image acquisition for all the slides in the fully automatic mode, it may be configured to re-acquire the image by resetting the imaging condition semi-automatically only for the image acquisition failure.

  The image acquisition device, the image acquisition method, and the image acquisition program according to the present invention are not limited to the above-described embodiments and configuration examples, and various modifications are possible. For example, in the above embodiment, the sample S is moved between the image acquisition position in the macro image acquisition unit 20 and the image acquisition position in the micro image acquisition unit 30 by the sample stage 15. Separate sample stages may be provided in the unit 20 and the micro image acquisition unit 30. In this case, the sample moving means is composed of only the sample transport unit 14. For the macro image acquisition unit 20, various configurations such as a configuration provided at a predetermined position of the sample transport unit 14 or a configuration provided in the sample storage unit 11 may be used.

  In the image acquisition device of the above embodiment, the macro image acquisition unit 20 and the micro image acquisition unit 30 are separately provided. However, the image acquisition optical can be switched according to the acquisition of the macro image or the micro image. By providing a system, the light source and the image acquisition unit may be configured as a single light source and a single image acquisition unit. In this case, examples of the switchable image acquisition optical system include an optical system including a revolver including a macro image acquisition objective lens and a micro image acquisition objective lens.

  In the above embodiment, the imaging device 31 in the micro image acquisition unit 30 is exemplified by an imaging device capable of acquiring a one-dimensional image and an imaging device capable of acquiring a two-dimensional image and TDI driving. For example, an imaging device such as a two-dimensional CCD camera (area sensor) may be used. In this case, as disclosed in US Pat. No. 6,816,606, a micro image of the entire sample S can be generated by acquiring and synthesizing a plurality of high resolution tile images.

  Moreover, in the said embodiment, although the image acquisition apparatus was comprised using the transmissive | pervious microscope system, it is not limited to this. For example, in the case of fluorescence observation, the light source may be epi-illumination instead of transmission illumination, and a reflection-type microscope system may be configured.

  The present invention can be used as an image acquisition device, an image acquisition method, and an image acquisition program that can efficiently execute image acquisition for each of a plurality of samples.

It is a block diagram which shows the structure of one Embodiment of an image acquisition apparatus. It is a figure which shows schematically the structure of the microscope part in an image acquisition apparatus. It is a figure which shows typically the acquisition method of the image of a sample. It is a graph which shows the setting method of the threshold value for binarizing a macro image. It is a block diagram which shows the structure of the control part in an image acquisition apparatus. It is a figure showing typically about creation of sample data using a micro picture. It is a flowchart shown about the image acquisition method in a fully automatic mode. It is a flowchart shown about the image acquisition method in semiautomatic mode. It is a flowchart shown about the image acquisition method in semiautomatic mode. It is a flowchart shown about the setting method of imaging conditions. It is a figure which shows the example of the operation screen used for the setting of an imaging condition. It is a figure which shows the example of the operation screen used for the setting of an imaging condition. It is a figure which shows the example of the operation screen used for the setting of an imaging condition.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 ... Microscope part, 11 ... Sample storage part, 12 ... Door, 13 ... Interlock mechanism, 14 ... Sample conveyance part, 15 ... Sample stage, 20 ... Macro image acquisition part, 21 ... Macro imaging device, 22 ... Imaging optics System: 25 ... Light source for macro, 30 ... Micro image acquisition unit, 31 ... Imaging device for micro, 32 ... Objective lens, 33 ... Z stage, 34 ... Light guide optical system, 35 ... Light source for micro, 36 ... Condensing lens , 38 ... Image correction unit, 40 ... Drive control unit, 41 ... Stage control unit, 42 ... Imaging device control unit, 43 ... Light source control unit,
DESCRIPTION OF SYMBOLS 60 ... Control part, 61 ... Display apparatus, 62 ... Input device, 70 ... Data processing part, 71 ... Imaging condition setting part, 72 ... Sample data preparation part, 73 ... Session management part, 75 ... Data storage part, 80 ... Image Acquisition control unit, 81... Macro image acquisition control unit, 82... Micro image acquisition control unit, 83.

Claims (16)

Sample storage means for storing a plurality of samples;
Macro image acquisition means for acquiring a macro image of the sample;
A micro image acquisition means for scanning the sample at a higher resolution than the macro image and acquiring the micro image;
Macro image acquisition control means for controlling the macro image acquisition operation by the macro image acquisition means, imaging condition setting means for setting the imaging condition of the micro image with reference to the macro image, and the micro image acquisition means Control means having a micro image acquisition control means for controlling the micro image acquisition operation,
The imaging condition setting means refers to the macro image, sets an image acquisition range and a focus measurement position as the imaging condition of the micro image,
The micro image acquisition control means acquires focus information for the sample based on the focus measurement position, and performs control to acquire the micro image based on the obtained focus information and the image acquisition range,
The control means has a semi-automatic mode as its control mode,
In the semi-automatic mode,
The macro image acquisition control unit is configured to acquire the macro image by disposing the sample at the storage position by the sample storage unit at the image acquisition position by the macro image acquisition unit for each of the plurality of samples. Performing control to place the sample in a standby position;
The imaging condition setting means sets the imaging condition of the micro image by allowing an operator to confirm the macro image and the imaging condition corresponding to the macro image for the sample from which the macro image has been acquired . After the macro image of the sample is acquired, in parallel with the acquisition of the macro image, setting the imaging conditions ,
The micro image acquisition control unit arranges the sample at the standby position and for which the imaging condition is set at the image acquisition position of the micro image acquisition unit, and acquires the micro image with reference to the imaging condition. performs control to,
In the semi-automatic mode,
The imaging condition setting means displays the imaging condition automatically obtained corresponding to the macro image on a display means, and asks an operator whether to use the imaging condition or to manually reset the imaging condition. An image acquisition apparatus characterized by being selected .   A sample moving means for moving each of the plurality of samples between the storage position, the standby position, and the image acquisition position in each of the macro image acquisition means and the micro image acquisition means; The image acquisition apparatus according to claim 1, wherein: The control means has a fully automatic mode as its control mode,
In the fully automatic mode,
The macro image acquisition control means performs control to acquire the macro image by arranging the sample in the storage position at the image acquisition position in the macro image acquisition means,
The imaging condition setting means automatically sets the imaging condition of the micro image corresponding to the macro image for the sample from which the macro image has been acquired,
The micro image acquisition control unit performs control for arranging the sample for which the imaging condition is set at an image acquisition position in the micro image acquisition unit and acquiring the micro image with reference to the imaging condition. The image acquisition apparatus according to claim 1 , wherein the image acquisition apparatus is an image acquisition apparatus. The macro image acquisition means and the micro image acquisition means are a single image acquisition means, and are configured to be capable of switching an image acquisition optical system in accordance with acquisition of the macro image or the micro image. image acquisition device according to any one of claims 1-3, characterized.   In the semi-automatic mode,
  The imaging condition setting unit displays an operation screen including a list display unit that displays a list of samples from which the macro image has been acquired together with a thumbnail image of the corresponding macro image on the display unit. 5. The image acquisition apparatus according to claim 1, wherein the image acquisition range and the focus measurement position which are the imaging conditions by automatic setting are displayed on the thumbnail image. 6.   In the semi-automatic mode,
  The imaging condition setting unit displays an operation screen including a list display unit that displays a list of samples from which the macro image has been acquired together with a thumbnail image of the corresponding macro image on the display unit. The image acquisition apparatus according to claim 1, wherein the imaging condition is set by magnifying and displaying the macro image of the selected sample on a macro image display unit. For multiple samples stored in the sample storage means,
A macro image acquisition step of acquiring a macro image of the sample;
Scanning the sample at a higher resolution than the macro image to obtain a micro image;
The macro image acquisition control step for controlling the macro image acquisition operation by the macro image acquisition step, the imaging condition setting step for setting the imaging condition of the micro image with reference to the macro image, and the micro image acquisition step A control step having a micro image acquisition control step for controlling a micro image acquisition operation,
The imaging condition setting step refers to the macro image, sets an image acquisition range and a focus measurement position as the imaging condition of the micro image,
The micro image acquisition control step acquires focus information for the sample based on the focus measurement position, and performs control to acquire the micro image based on the obtained focus information and the image acquisition range,
The control step has a semi-automatic mode as its control mode,
In the semi-automatic mode,
In the macro image acquisition control step, for each of the plurality of samples, the sample in the storage position by the sample storage means is arranged at the image acquisition position in the macro image acquisition step, and the macro image is acquired. Performing control to place the sample in a standby position;
The imaging condition setting step sets an imaging condition of the micro image by allowing an operator to confirm the macro image and the imaging condition corresponding to the macro image for the sample from which the macro image has been acquired . After the macro image of the sample is acquired, in parallel with the acquisition of the macro image, setting the imaging conditions ,
In the micro image acquisition control step, the sample in which the imaging condition is set at the standby position is arranged at the image acquisition position in the micro image acquisition step, and the micro image is acquired with reference to the imaging condition. performs control to,
In the semi-automatic mode,
In the imaging condition setting step, the imaging condition automatically obtained corresponding to the macro image is displayed on the display means, and the operator is asked whether to use the imaging condition or to manually reset the imaging condition. An image acquisition method characterized by selecting .   A sample moving step of moving each of the plurality of samples between the storage position, the standby position, and the image acquisition position in each of the macro image acquisition step and the micro image acquisition step. The image acquisition method according to claim 7, wherein: The control step has a fully automatic mode as its control mode,
In the fully automatic mode,
The macro image acquisition control step performs control to acquire the macro image by arranging the sample in the storage position at the image acquisition position in the macro image acquisition step,
The imaging condition setting step automatically sets the imaging condition of the micro image corresponding to the macro image for the sample from which the macro image is acquired,
In the micro image acquisition control step, the sample in which the imaging condition is set is arranged at an image acquisition position in the micro image acquisition step, and control is performed to acquire the micro image with reference to the imaging condition. The image acquisition method according to claim 7 or 8, characterized in that   In the semi-automatic mode,
  In the imaging condition setting step, an operation screen including a list display unit that displays a list of samples from which the macro image is acquired together with a thumbnail image of the corresponding macro image is displayed on the display unit. The image acquisition method according to any one of claims 7 to 9, wherein the image acquisition range and the focus measurement position which are the imaging conditions by automatic setting are displayed on the thumbnail image.   In the semi-automatic mode,
  In the imaging condition setting step, an operation screen including a list display unit for displaying a list of samples from which the macro image is acquired together with a thumbnail image of the corresponding macro image is displayed on the display means, The image acquisition method according to any one of claims 7 to 10, wherein the imaging condition is set by enlarging and displaying the macro image of the selected sample on a macro image display unit. Sample storage means for storing a plurality of samples;
Macro image acquisition means for acquiring a macro image of the sample;
The sample is scanned at a higher resolution than the macro image, and is applied to an image acquisition device including a micro image acquisition means for acquiring the micro image,
Macro image acquisition control processing for controlling the macro image acquisition operation by the macro image acquisition means, imaging condition setting processing for setting the imaging conditions of the micro image with reference to the macro image, and the micro image acquisition means by the micro image acquisition means Including a control process having a micro image acquisition control process for controlling a micro image acquisition operation;
The imaging condition setting process refers to the macro image, sets an image acquisition range and a focus measurement position as the imaging condition of the micro image,
The micro image acquisition control process acquires focus information for the sample based on the focus measurement position, and performs control to acquire the micro image based on the obtained focus information and the image acquisition range,
The control process has a semi-automatic mode as its control mode,
In the semi-automatic mode,
The macro image acquisition control process, for each of the plurality of samples, arranges the sample at the storage position by the sample storage means at the image acquisition position by the macro image acquisition means, and acquires the macro image, Performing control to place the sample in a standby position;
The imaging condition setting process sets the imaging condition of the micro image by letting an operator confirm the macro image and the imaging condition corresponding to the macro image for the sample from which the macro image has been acquired . After the macro image of the sample is acquired, in parallel with the acquisition of the macro image, setting the imaging conditions ,
In the micro image acquisition control process, the sample in which the imaging condition is set at the standby position is arranged at an image acquisition position in the micro image acquisition unit, and the micro image is acquired with reference to the imaging condition. performs control to,
In the semi-automatic mode,
In the imaging condition setting process, the imaging condition automatically obtained corresponding to the macro image is displayed on a display unit, and the operator is asked whether to use the imaging condition or to manually reset the imaging condition. An image acquisition program for causing a computer to execute the control process to be selected .   The computer executes a sample movement process for moving each of the plurality of samples between the storage position, the standby position, and the image acquisition position in each of the macro image acquisition unit and the micro image acquisition unit. The image acquisition program according to claim 12. The control process has a fully automatic mode as its control mode,
In the fully automatic mode,
The macro image acquisition control process performs control to acquire the macro image by arranging the sample in the storage position at the image acquisition position in the macro image acquisition means,
The imaging condition setting process automatically sets the imaging condition of the micro image corresponding to the macro image for the sample from which the macro image is acquired,
The control for performing the control for acquiring the micro image with reference to the imaging condition by arranging the sample for which the imaging condition has been set at an image acquisition position in the micro image acquisition unit in the micro image acquisition control process The image acquisition program according to claim 12 or 13, which causes a computer to execute processing.   In the semi-automatic mode,
  In the imaging condition setting process, an operation screen including a list display unit that displays a list of samples from which the macro image is acquired together with a thumbnail image of the corresponding macro image is displayed on the display unit. The image acquisition program according to any one of claims 12 to 14, wherein the image acquisition range and the focus measurement position which are the imaging conditions by automatic setting are displayed on the thumbnail image.   In the semi-automatic mode,
  The imaging condition setting process displays an operation screen including a list display unit that displays a list of samples from which the macro image has been acquired together with a thumbnail image of the corresponding macro image on the operation screen. The image acquisition program according to any one of claims 12 to 15, wherein the imaging condition is set by enlarging and displaying the macro image of the selected sample on a macro image display unit.

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